شناسایی مولکولی گونه های آفت Heliothine (Lep. Noctuidae) با مشخصات PCR-RFLP هسته‌ای و میتوکندریایی

نوع مقاله : مقاله کامل، انگلیسی

نویسندگان

1 مرکز دانشگاهی برزیل، برازیلیا، برزیل

2 گروه علوم فیزیولوژیکی، موسسه علوم زیستی، دانشگاه برازیلیا، برازیلیا، برزیل

3 اداره دولتی آموزش. معاون برنامه ریزی، نظارت و ارزیابی آموزشی، سوپلاو، برزیل

4 اتحادیه تحقیقات کشاورزی برزیل (Embrapa)، بخش منابع ژنتیکی و بیوتکنولوژی – آزمایشگاه باکتری های بیمارگر حشرات، برازیلیا ، برزیل

10.61186/jesi.44.4.10

چکیده

آرایه­ بندی زیرخانواده Heliothinae پیچیده است، به‌ویژه شناسایی این گونه‌ها در مرحله لاروی با مانع مواجه می­شود. روش جایگزین، استفاده از روش‌های مولکولی مانند تکنیک PCR-RFLP (یا پلی‌مورفیسم طول قطعه محدود) است که به کمک و تکمیل شناسایی برخی از گونه‌های Heliothinae کمک می‌کند. هدف از این مطالعه ایجاد مشخصات PCR-RFLP با استفاده از نشانگرهای مولکولی برای شناسایی چهار گونه Heliothinae بود. آغازگرهای اختصاصی برای تکثیر ژن‌های سیتوکروم اکسیداز زیر واحد یک (COI) و  ژن EF-1a برای شناسایی گونه‌های Helicoverpa armigera، H. zea، H. gelotopoeon و Chloridea virescens ساخته شدند. در هضم ژن COI با آنزیم BfaI، قطعات 294 جفت باز و 321 جفت باز برای H. armigera به دست آمد، در حالی که برای H. zea، H. gelotopoeon و C. virescens هیچ نقطه برشی وجود نداشت. آنزیم HpaI قطعات 58 جفت باز و 557 جفت باز برای H. gelotopoeon ایجاد نمود، در حالی که  برای  گونه های H. armigera، H. zea و C. virescens هیچ گونه برشی رخ نداد.  برش ناحیه ژنی EF-1a با آنزیم EcoRV  برای C. virescens دو قطعه به طول 142 و 964 باز ایجاد کرد، در حالی که برای سایر گونه­ ها هیچ  نقطه برشی نداشت. مارکر مولکولی مبتنی بر برش آنزیمی نواحی ژنی مذکور می توانند به حشره شناسان برای تمایز بین گونه‌های H. zea، H. armigera، H. gelotopoeon و C. virescens کمک کنند و همچنین می­ توانند به عنوان ابزاری برای نظارت بر  محدوده پراکنش این آفات در مناطق کشاورزی استفاده شوند.

چکیده تصویری

شناسایی مولکولی گونه های آفت Heliothine (Lep. Noctuidae) با مشخصات PCR-RFLP هسته‌ای و میتوکندریایی

کلیدواژه‌ها

موضوعات

عنوان مقاله [English]

Molecular identification of Heliothine species by nuclear and mitochondrial PCR-RFLP profile

نویسندگان [English]

  • Paulo Queiroz 1
  • Elias Ferreira Sabiá Júnior 2
  • Erica Martins 3
  • Rose Monnerat 4
1 University Center of Brasília - CEUB - SEPN, 707/907, Via W 5 Norte, 70790-075, Asa Norte/Brasília (Distrito Federal), Brazil
2 Department of Physiological Sciences, Institute of Biological Sciences, University of Brasília - UnB, 70910-900, Brasília (Distrito Federal), Brazil.
3 State Department of Education – SEE – DF. Undersecretary of Planning, Monitoring and Educational Assessment – SUPLAV. SBN Quadra 02 Bloco C, Edifício Phenícia, 11º andar, 70.040-020, Brasília (Distrito Federal), Brazil
4 Embrapa Genetic Resources and Biotechnology – Laboratory of Entomopathogenic Bacteria. Parque Estação Biológica, PqEB, Av. W5 Norte (final) CP 02372, 70770-917, Brasília (Distrito Federal), Brazil
چکیده [English]

The taxonomy of the Heliothinae subfamily is complex, often becoming an obstacle in the identification of these species, especially during the larval stage. An alternative is the use of molecular methods such as the PCR-RFLP technique (or Restriction Fragment Length Polymorphism), which helps to aid and complement the identification of some heliothine species. The aim of this study was to establish the PCR-RFLP profile using molecular markers for the identification of four heliothine species. Specific primers were constructed for the amplification of the cytochrome oxidase I (COI) and elongation factor 1 alpha (EF-1) genes for identifying Helicoverpa armigera, H. zea, H. gelotopoeon, and Chloridea virescens species. In the digestion of the COI gene with the BfaI enzyme, fragments of 294 bp and 321 bp were obtained for H. armigera, while there was no digestion for H. zea, H. gelotopoeon, and C. virescens. The HpaI enzyme produced fragments of 58 bp and 557 bp for H. gelotopoeon, while there was no digestion for H. armigera, H. zea, and C. virescens. The digestion of the EF-1 gene with the EcoRV enzyme produced fragments of 142 bp and 964 bp for C. virescens, while there was no cleavage for the other heliothines. These molecular markers can help the entomologists, aiding in the discrimination between H. zea, H. armigera, H. gelotopoeon, and C. virescens, and can also be used as a tool for monitoring the spread of these pests in agricultural regions.

کلیدواژه‌ها [English]

  • molecular entomology, Commodities
  • Cytochrome oxidase I
  • Helicoverpa
  • Nuclear elongation factor 1 alpha

© 2024 by Author(s), Published by the Entomological Society of Iran

This Work is Licensed under Creative Commons Attribution-Non Commercial 4.0 International Public License

مراجع

Albernaz, K. C., Czepak, C., Costa, J., Zuntini, B. & Borges, M. (2014) Guia de Identificação Helicoverpa armigera. [Online] Available at: http://www.plantaconsultoria.com/helicoverpa-armigera-guia-de-identificacao/.
Arneodo, J. D., Balbi, E. I., Flores, F. M. & Sciocco-Cap, A. (2015) Molecular identification of Helicoverpa armigera (Lepidoptera: Noctuidae: Heliothinae) in Argentina and development of a novel PCR-RFLP method for its rapid differentiation from H. zea and H. gelotopoeon. Journal of Economic Entomology 108(6), 2505-2510. doi: 10.1093/jee/tov254.
Ávila, C. J., Vivan, L. M. & Tomquelski, G. V. (2013) Ocorrência, aspectos biológicos, danos e estratégias de manejo de Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) nos sistemas de produção agrícolas. Circular Técnica 23, 12p.
Avise, J. C. (2004). Molecular markers, natural history, and evolution, 2nd ed. Sinauer Associates, Sunderland, MA.
Behere, G. T., Tay, W. T., Russell, D. A., Heckel, D. G., Appleton, B. R., Kranthi, K. R. & Batterham, P. (2007) Mitochondrial DNA analysis of field populations of Helicoverpa armigera (Lepidoptera: Noctuidae) and of its relationship to H. zea. BMC Ecology and Evolution. 14(7), 117. doi: 10.1186/1471-2148-7-117.
Behere, G. T., Tay, W. T., Russell, D. A. & Batterham, P. (2008) Molecular markers to discriminate among four pest species of Helicoverpa (Lepidoptera: Noctuidae). Bulletin of Entomological Research 98, 599–603. doi: 10.1017/S0007485308005956.
Bueno, R. C. O. F., Yamamoto, P. T., Carvalho, M. M. & Bueno, N. M. (2014) Occurrence of Helicoverpa armigera (Hübner, 1808) on citrus in the state of Sao Paulo, Brazil. Revista Brasileira de Fruticultura 36(2), 520-523. https://doi.org/10.1590/0100-2945-357/13.
Che, L. H., Zhang, S. Q., Li, Y., Liang, D., Pang, H., Ślipiński, A. & Zhang, P. (2017) Genome-wide survey of nuclear protein-coding markers for beetle phylogenetics and their application in resolving both deep and shallow-level divergences. Molecular Ecology Resours. 17(6), 1342-1358. doi: 10.1111/1755-0998.12664.
Czepak, C., Albernaz, K. C., Vivan, L. M., Guimarães, H. O. & Carvalhais, T. (2013) Primeiro registro de ocorrência de Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) no Brasil. Pesquisa Agropecuária Tropical 43, 110–113. https://doi.org/10.1590/S1983-40632013000100015.
Daly, J. C. & Gregg, P. (1985) Genetic variation in Heliothis in Australia: species identification and gene flow in the two pest species H. armigera (Hübner) and H. punctigera (Wallengren) (Lepidoptera: Noctuidae). Bulletin of Entomological Research 75, 169–184. https://doi.org/10.1017/S0007485300014243.
Dean, A. N., Niemi, J. B., Tyndall, J. C., Hodgson, E. W., O'Neal, M. E. (2021) Developing a decision-making framework for insect pest management: a case study using Aphis glycines (Hemiptera: Aphididae). Pest Management Science 77(2), 886-894. doi: 10.1002/ps.6093.
Ferrelli, M. L., Taibo, C., Fichetti, P., Sciocco-Cap, A. & Arneodo, J. D. (2016) Characterization of a new Helicoverpa armigera nucleopolyhedrovirus variant causing epizootic on a previously unreported host, Helicoverpa gelotopoeon (Lepidoptera: Noctuidae). Journal of Invertebrate Pathology 138, 89-93. doi: 10.1016/j.jip.2015.08.009.
Kearse, M., Moir, R., Wilson, A., Stones-Havas, S., Cheung, M., Sturrock, S., Buxton, S., Cooper, A., Markowitz, S., Duran, C., Thierer, T., Ashton, B., Mentjies, P., Drummond, A. (2012) Geneious Basic: An integrated and extendable desktop software platform for the organization and analysis of sequence data. Bioinformatics 28(12), 1647-1649. doi: 10.1093/bioinformatics/bts199.
Koressaar, T. & Remm, M. (2007) Enhancements and modifications of primer design program Primer3. Bioinformatics 23(10), 1289-91. doi: 10.1093/bioinformatics/btm091.
Kranthi, S., Kranthi, K. R., Bharose, A. A. & Syed, S. N. (2005) A PCR-RFLP tool for differentiating Helicoverpa armigera and H. assulta (Lepidoptera: Noctuidae). Current Science 89, 1322–1323. https://doi.org/10.1093/jee/tov254.
Krishnarao, G., Sujatha, A., Sudhagar, S., Raghava, Thimmappa, Suneetha, P., Gundappa, B., Singh, H. S., Vyas, M., & Damodaram, K. J. P. (2019) Molecular characteristics of Indian population of Deanolis sublimbalis Snellen (Crambidae: Lepidoptera) using mitochondrial and nuclear genes. Oriental Insects 54(1), 16. doi: 10.1080/00305316.2019.1673257.
Lammers, J. W. & MacLeod, A. (2007) Report of a pest risk analysis: Helicoverpa armigera (Hübner, 1808). [Online] Available at:  http://www.fera.defra.gov.uk/plants/plantHealth/pestsDiseases/documents/helicoverpa.pdf.
Leite, L. A. R., Casagrande, M. M. & Mielke, O. H. H. (2013) External morphology of the adult of Dynamine postverta (Cramer) (Lepidoptera, Nymphalidae, Biblidinae) and patterns of morphological similarity among species from eight tribes of Nymphalidae. Revista Brasileira de Entomologia 57, 133‑148. https://doi.org/10.1590/S0085-56262013005000006.
Mastrangelo, T., Paulo, D. F., Bergamo, L. W., Morais, E. G. F., Silva, M., Bezerra-Silva, G. & Azeredo-Espin, A. M. L. (2014) Detection and Genetic Diversity of a Heliothine Invader (Lepidoptera: Noctuidae) From North and Northeast of Brazil. Journal of Economic Entomology 107(3), 970-80. doi: 10.1603/ec13403.
Ming, Q.  L. & Wang, C. Z. (2006) Genetic differentiation of Helicoverpa armigera (Hübner) and H. assulta (Guenée) (Lepidoptera: Noctuidae) based on AFLP markers. Insect Science 13, 437–444. https://doi.org/10.1111/j.1744-7917.2006.00113.x.
Mitchell, A. & Gopurenko, D. (2016) DNA Barcoding the Heliothinae (Lepidoptera: Noctuidae) of Australia and Utility of DNA Barcodes for Pest Identification in Helicoverpa and Relatives. PLOS One 11(8), e0160895. doi: 10.1371/journal.pone.0160895.
Monnerat, R., Martins, E., Queiroz, P., Ordúz, S., Jaramillo, G., Benintende, G., Cozzi, J., Real, M. D, Martinez-Ramirez, A., Rausell, C., Cerón, J., Ibarra, J. E., Del Rincon-Castro, M. C., Espinoza, A. M., Meza-Basso, L., Cabrera, L., Sánchez, J., Soberon, M. &  Bravo, A. (2006) Genetic variability of Spodoptera frugiperda Smith (Lepidoptera: Noctuidae) populations from Latin America is associated with variations in susceptibility to Bacillus thuringiensis cry toxins. Applied and Environmental Microbiology 72(11): 7029-35. doi: 10.1128/AEM.01454-06.
Murúa, M. G., Scalora, F. S., Navarro, F. R., Cazado, L. E. Casmuz, A., Villagrán, M. E., Lobos, E. & Gastaminza, G. (2014) First record of Helicoverpa armigera (Lepidoptera: Noctuidae) in Argentina. Florida Entomologist 97, 854–856. https://doi.org/10.1653/024.097.0279.
Pearce, S. (2003) The use of naturally occurring arthropod predators for the control of Helicoverpa spp. in grain crops in southeast Queensland. PhD thesis, The University of Queensland, Brisbane, Australia.
Pedgley, D. E. (1985) Windborne migration of Heliothis armigera (Hübner) (Lepidoptera: Noctuidae) to the British Isles. Entomologist's Gazette, 36(1), 15-20.
Pogue, M. G. (2004) A new synonym of Helicoverpa zea (Boddie) and differentiation of adult males of H. zea and H. armigera (Hübner) (Lepidoptera: Noctuidae: Heliothinae). Annals of the Entomological Society of America 97(6), 1222-1226. https://doi.org/10.1603/0013-8746(2004)097[1222:ANSOHZ]2.0.CO;2.
Pogue, M. G. (2013) Revised status of Chloridea Duncan and (Westwood), 1841, for the Heliothis virescens species group (Lepidoptera: Noctuidae: Heliothinae) based on morphology and three genes. Systematic Entomology 38. doi:10.1111/syen.12010.
Specht, A., Sosa-Gómez, D. N., Paula-Moraes, S. V. & Yano, S. A. C. (2013) Identificação morfológica e molecular de Helicoverpa armigera (Lepidoptera: Noctuidae) e ampliação de seu registro de ocorrência no Brasil. Pesquisa Agropecuária Brasileira 48(6), 689-692. https://doi.org/10.1590/S0100-204X2013000600015.
Tay, W. T., Soria, M. F., Walsh, T., Thomazoni, D., Silvie, P., Behere, G. T., Anderson, C. & Downes, S. (2013) A brave new world for an old-world pest: Helicoverpa armigera (Lepidoptera: Noctuidae) in Brazil. PLOS One, 8(11), e80134. https://doi.org/10.1371/journal. pone.0080134.
Trowell, S. C., Forrester, N. W., Garsia, K. A., Lang, G. A., Bird, L. J., Hill, A. S., Skerritt, J. H. & Daly, J. (2000) Rapid antibody-based field test to distinguish between Helicoverpa armigera (Lepidoptera: Noctuidae) and Helicoverpa punctigera (Lepidoptera: Noctuidae). Journal of Economic Entomology 93, 878–891. https://doi.org/10.1603/0022-0493-93.3.878,
Tsai, C. L., Chu, I. H., Chou, M. H., Chareonviriyaphap, T., Chiangm M. Y., Lin, P. A., Lu, K. H. & Yeh, W. B. (2020) Rapid identification of the invasive fall armyworm Spodoptera frugiperda (Lepidoptera, Noctuidae) using species-specific primers in multiplex PCR. Scientific Reports 10(1), 16508. doi: 10.1038/s41598-020-73786-7.
Untergrasser, A., Cutcutache, I., Koressaar, T. Ye. J., Faircloth, B. C., Remm, M. & Rozen, S. G. (2012) Primer3 - new capabilities and interfaces. Nucleic Acids Research 40(15), e115. doi: 10.1093/nar/gks596.
Vincze, T., Posfai, J. & Roberts, R. J. (2003) NEBcutter: a program to cleave DNA with restriction enzymes. Nucleic Acids Research 31, 3688-3691. https://doi.org/10.1093/nar/gkg526.
Walsh, T. K., Perera, O., Anderson, C., Gordon, K., Czepak, C., McGaughran, A., Zwick, A., Hackett, D. & Tay, W. T. (2019) Mitochondrial DNA genomes of five major Helicoverpa pest species from the Old and New Worlds (Lepidoptera: Noctuidae). Ecology and Evolution 9(5), 2933-2944. doi: 10.1002/ece3.497.

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